Enzymatic template polymerization

a template and enzyme technology, applied in the field of enzyme-based template polymerization, can solve the problems of limited application possibilities of polymers, and achieve the effects of limiting parasitic branching, improving electrical and optical properties of the final polymer complex, and stabilizing desired electronic and optical properties

Inactive Publication Date: 2005-04-21
UNITED STATES OF AMERICA THE AS REPRESENTED BY THE SEC OF THE ARMY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Benefits of technology

[0006] Another aspect of the invention is a method of modulating the conformation of a polynucleotide double helix which is bound to a conductive polymer as a complex by changing the oxidation state of the conductive polymer. In a specific embodiment, polyaniline is bound to a polynucleotide double helix as a complex. Oxidation of polyaniline (e.g., increasing the positive charge on the polyaniline) which is complexed to a polynucleotide double helix causes the double helix to become more tightly wound (i.e., the double helix will have more base pairs per turn after oxidation of the polyaniline). Conversely, reducing the polyaniline will cause a double helix associated with it to become more loosely wound. Therefore, complexation of polyaniline to a polynucleotide double helix provides a method of modulating the conformation of the double helix by changing the oxidation state of the polyaniline.
[0010] In invention, the polynucleotide can serve at least three critical functions. First, the polynucleotide can serve as a template upon which the monomers preferentially align themselves to form a complex, such as a charge-transfer complex, thereby limiting parasitic branching and controlling the shape of the polymer. In the case of polyaniline, the mechanism of polymerization is primarily para-directed and results in formation of the electrically active form of polyaniline. This preferential alignment provides improved electrical and optical properties of the final polymer complex. Second, the polynucleotide can serve as a large molecular dopant species which is complexed and essentially locked to the polyaniline chains. Current limitations to the actual use of polyaniline in electronic and optical applications largely has been due to poor dopant stability. Small ionic dopants or chromophores that are currently used are known to diffuse away with time and / or conditions. Locking of a large polyelectrolyte dopant (e.g., a polynucleotide) to the polymer is significant in that it ensures that the electronic nature of the conjugated backbone structure of the polymer is maintained, and hence the desired electronic and optical properties are stabilized. Third, the polynucleotide template can improve water solubility of the final polynucleotide / polyaniline complex for environmentally friendly, facile, and inexpensive processing.

Problems solved by technology

However, the potential applications to which polymers can be put have been limited by their lack of solubility and processability.

Method used

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  • Enzymatic template polymerization
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Examples

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example 1

[0068] A. Materials and Methods

[0069] Horseradish peroxidase (HRP) (enzyme classification number (EC) 1.11.1.7), phosphate and Tris-HCl buffers were obtained from Sigma Chemicals Company, St. Louis, Mo. Aniline, sulfonated polystyrene (SPS) and hydrogen peroxide (30%) were obtained from Aldrich Chemicals, Inc., Milwaukee, Wis. All the chemicals were used as received.

[0070] B. Results and Discussion

[0071] The progress of a template-assisted polymerization reaction of aniline in the presence of the polyelectrolyte, sulfonated polystyrene (SPS) in a 1:1 ratio, was monitored by the change in visible absorbance. A Perkin-Elmer Lambda-9® UV-Vis-near IR spectrophotometer was used for the spectral characterization of the polymer. FIG. 3 shows the visible absorption spectra of the sulfonated polystyrene / polyaniline (SPS / PA) complex prepared under various pH conditions of 4, 6, 8, and 10. As shown in FIG. 3, SPS / PA, prepared at a pH of 4, exhibited a strong absorbance maximum at approximat...

example 2

[0077] A. Materials and Methods

[0078] Horseradish peroxidase (HRP) (enzyme classification number (EC) 1.11.1.7), phosphate and Tris-HCl buffers were obtained from Sigma Chemicals Company, St. Louis, Mo. Phenol, sulfonated polystyrene (SPS) and hydrogen peroxide (30%) were obtained from Aldrich Chemicals, Inc., Milwaukee, Wis. All the chemicals were used as received.

[0079] B. Results and Discussion

[0080] The progress of a template-assisted polymerization reaction of phenol in the presence of the polyelectrolyte, sulfonated polystyrene (SPS) in a 1:1 ratio, was monitored by the change in visible absorbance. Perkin-Elmer Lambda-9® UV-Vis-near IR spectrophotometer was used for the spectral characterization of the polymer. FIG. 7A shows the visible absorption of polyphenol without SPS, versus phenol monomer. As shown, there was a significant absorption maximum in the visible spectrum upon polymerization, indicating formation of polyphenol. However, with time the polymer began to preci...

example 3

[0082] A. Preparation of DNA-Polyaniline Complex

[0083] Horseradish peroxidase (HRP, EC 1.11.1.7) type II, (150-200 units / mg) solid was purchased from Sigma Chemical Co. (St. Louis, Mo.). Calf Thymus DNA was purchased from Worthington Biochemical Corporation (Freehold, N.J.). Aniline monomer (purity 99.5%) and hydrogen peroxide (30% by weight) were purchased from Aldrich Chemicals, Inc., Milwaukee, Wis., and were used as received. All other chemicals were of reagent grade or better. All glassware and plasticware were sterilized by autoclaving for 30 minutes.

[0084] A stock solution of calf Thymus DNA (19.9 mg) was dissolved in 40 ml of sterile, 10 mM sodium citrate buffer maintained at pH 4. The solution was stored in the refrigerator for 48 hours before reaction. The concentration of DNA was determined by the UV absorbance at 258 nm. The molar extinction coefficient (ε) at 258 nm was taken as 6420 1 mol−1 cm1, as reported by Sprecher, et al., Biopolymers (1979), 18: 1009. The react...

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Abstract

A conductive polymer is formed enzymatically in the presence of a polynucleotide template. The method includes combining at least one redox monomer with a polynucleotide template and a redox enzyme, such as horseradish peroxidase, to form a reaction mixture. The monomer aligns along the template before or during the polymerization. Therefore, the polynucleotide template thereby affects the molecular weight and conformation of the conductive polymer. When the conductive polymer is complexed to a polynucleotide duplex, the conformation of the polynucleotide duplex can be modulated by changing the oxidation state of the conductive polymer.

Description

RELATED APPLICATION [0001] This is a continuation of U.S. application Ser. No. 09 / 447,987, filed Nov. 23, 1999, which is a continuation-in-part of U.S. application Ser. No. 08 / 999,542, filed Nov. 21, 1997 (now U.S. Pat. No. 6,018,018), which is a continuation-in-part of U.S. application Ser. No. 08 / 915,827, filed Aug. 21, 1997 (now U.S. Pat. No. 5,994,498), the entire teachings of which are incorporated herein by reference.GOVERNMENT SUPPORT [0002] This invention was made with support from the Government under ARO Cooperative Grant DAAH04-94-2-003. The Government has certain rights in this invention.BACKGROUND OF THE INVENTION [0003] Recently, there has been an increased interest in tailored development of certain classes of polymers, such as electrically conductive and optically active polymers (e.g. polythiophene, polypyrrole, polyphenols and polyaniline) for application to wider ranges of use. Examples of such uses include light-weight energy storage devices, electrolytic capacit...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C08G73/02H01B1/12
CPCC08G73/0266Y10T436/143333Y10S977/704H01B1/128
Inventor SAMUELSON, LYNNE A.BRUNO, FERDINANDOTRIPATHY, SUKANT K.TRIPATHY, SUSANNAGARAJAN, RAMASWAMYKUMAR, JAYANTLIU, WEI
Owner UNITED STATES OF AMERICA THE AS REPRESENTED BY THE SEC OF THE ARMY
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